黃大野+姚經武+朱志剛等
摘要:研究枯草芽孢桿菌(Bacillus subtilis)水分散粒劑防治黃瓜立枯病效果及對黃瓜的促生作用。盆栽試驗表明,1012 CFU/g 枯草芽孢桿菌水分散粒劑對黃瓜立枯病具有良好的防效并具有一定的促生作用。在濃度為2 000 μg/mL和1 000 μg/mL時,對黃瓜立枯萎病的防效為71.48%和52.02%。在濃度為2 000 μg/mL時,黃瓜發芽率、株高、根長和鮮重分別較對照提高了10.57%、28.58%、29.63%和27.46%。
關鍵詞:枯草芽孢桿菌(Bacillus subtilis);水分散粒劑;黃瓜立枯病;促生作用
中圖分類號:S436.5 文獻標識碼:A 文章編號:0439-8114(2014)18-4327-02
立枯絲核菌(Rhizoctonia solani)是一種毀滅性的土傳病害病原菌,能侵染多種大田和園藝作物,如棉花、水稻和黃瓜等,并引起嚴重減產[1-3]。目前預防由立枯絲核菌引起的病害主要依靠栽培技術[4],當危害特別嚴重時,往往要使用化學殺菌劑[5]。然而,目前的栽培技術和化學防治不能完全有效控制病害的發生,并且化學藥劑的使用會造成環境污染。
生物防治是防治立枯病的一種有效和環境友好的方法。多種微生物,如深綠木霉(Trichoderma atroviride)、哈慈木霉(Trichoderma harzianum)、熒光假單胞桿菌(Pseudomonas fluorescens)和枯草芽孢桿菌(Bacillus subtilis)已經被證明能有效防治由立枯絲核菌引起的植物病害,其中,應用最多的是枯草芽孢桿菌[6-9]。目前,國內外已經有多種優良性狀的野生菌株登記用于防治多種作物的葉部和土傳病害,生產上所使用的劑型多為可濕性粉劑,本研究利用自主研發的枯草芽孢桿菌環保型新劑型水分散粒劑防治黃瓜立枯病,為枯草芽孢桿菌水分散粒劑推廣應用于防治黃瓜立枯病奠定基礎。
1 材料與方法
1.1 材料
病原菌:立枯絲核菌,由湖北省生物農藥工程研究中心分離保存。
供試藥劑:1012 CFU/g枯草芽孢桿菌水分散粒劑,由湖北省生物農藥工程研究中心研制;枯草芽孢桿菌可濕性粉劑Serenade,由美國AgraQuest公司提供,50%多菌靈可濕性粉劑,由江蘇百靈農化股份有限公司生產。
供試植物:黃瓜,品種為中農8號
1.2 方法
1.2.1 枯草芽孢桿菌水分散粒劑防治黃瓜立枯病試驗 將立枯絲核菌從保存斜面轉到PDA培養基進行活化,27 ℃培養5 d, 隨后將其加入含有3%風干燕麥粉的砂土進行混合,接種的混合物在27 ℃和20%濕度條件下培養21 d,然后將培養物以3∶100比例與草炭土混合[10],成為帶菌土,用于播種黃瓜。
將黃瓜種子浸泡于3%次氯酸鈉水溶液中10 min,然后用無菌水反復沖洗5次。隨后將黃瓜種子放于墊有紗布的培養皿中,28 ℃催芽48 h。將發芽種子分別浸種于2 000 μg/mL 枯草芽孢桿菌水分散粒劑、1 000 μg/mL 枯草芽孢桿菌水分散粒劑、500 μg/mL 多菌靈和無菌水中2 h,然后播種于裝有帶菌土的穴盤中,溫室條件下常規管理,播種10 d后觀察幼苗發病情況并統計病株數。每個處理3次重復,每個重復20株苗。
1.2.2 枯草芽孢桿菌對黃瓜的促生作用試驗 對黃瓜種子進行表面消毒,方法同“1.2.1”,將消毒種子分別浸種于2 000 μg/mL枯草芽孢桿菌水分散粒劑、2 000 μg/mL Serenade和無菌水中,25 ℃浸種24 h后催芽并統計各處理發芽率。每個處理3次重復,每個重復100粒種子。將每個處理芽長一致的種子播種于育苗缽中,溫室常規管理,在試驗過程中不添加任何營養物質。20 d后測定株高、根長、鮮重,每個處理3次重復,每個重復20株苗。計算種子活力,種子活力=發芽率×(平均根長+平均株高)[11]。
2 結果與分析
盆栽試驗結果表明,枯草芽孢桿菌水分散粒劑浸種對黃瓜立枯病具有良好的防治效果(表1),在2 000 μg/mL條件下浸種對黃瓜立枯病的防效達到71.48%,1 000 μg/mL時防效也達到50%以上,防效低于對照化學藥劑多菌靈。
枯草芽孢桿菌水分散粒劑對黃瓜具有明顯的促生作用(表2),2 000 μg/mL浸種處理,種子發芽率從82.00%提高到90.67%,發芽率、株高、根長和鮮重分別較對照提高了10.57%、28.58%、29.63%和27.46%,并且種子活力有了明顯的提高,促生作用與商品化的枯草芽孢桿菌制劑Serenade相當。
3 討論
枯草芽孢桿菌是一種低毒微生物殺菌劑,對多種作物表現出良好的防病與促生作用[12]。但目前生產上常用的劑型主要是10億活芽孢/g可濕性粉劑、1 000億活孢子/g可濕性粉劑,劑型較為單一。可濕性粉劑配制噴灑液時容易發生藥粉飛揚,對操作人員造成危害,包裝袋內殘剩的藥粉污染環境,而水分散粒劑則是最具發展前景的劑型之一,可以避免可濕性粉劑中粉塵對操作者的毒害以及對環境的污染,并且性能優異,生物活性較高[13]。
本研究中的枯草芽孢桿菌水分散粒劑是一種較為理想的微生物殺菌劑劑型,其在活體條件下對黃瓜立枯病表現出良好的防病與促生作用,其田間防治效果還有待進一步的研究。同時,對其他病害,特別是葉部病害的防治效果需要進一步的研究,為其進一步的推廣應用奠定基礎。
參考文獻:
[1] OU S H. Rice disease[M]. AUS Kew: the Cambrian News Ltd,1985.
[2] OGOSHI A. Ecology and pathogenicity of anastomosis and intraspecific groups of Rhizoctonia solani Kühn[J]. Annual Review of Phytopathology,1987,25:125-143.
[3] STRASHNOV Y, ELAD Y, SIVAN A, et al. Control of Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai [J]. Crop Protection,1985,4:359-364.
[4] SECOR G A, GUDMESTAD NC. Managing fungal diseases of potato[J]. Canadian Journal of Plant Pathology, 1999, 21: 213-221.
[5] Parry D W. Diseases of potato[A]. Plant pathology in agriculture [M]. UK: Cambridge University Press,1981.
[6] REITHNER B, SCHUHMACHER R, STOPPACHER N, et al. Signaling via the Trichoderma atroviride mitogen-activated protein kinase Tmk1 differentially affects mycoparasitism and plant protection[J]. Fungal Genetics and Biology:FG&B,2007,44(11): 1123-1133.
[7] HADAR Y, CHET I, HENIS Y. Biological control of Rhizoctonia solani damping-off with wheat bran culture of Trichoderma harzianum[J]. Phytopathology,1979,69:64-68.
[8] NAGARAJKUMAR M, BHASKARAN R, VELAZHAHAN R. Involvement of secondary metabolites and extracellular lytic enzymes produced by Pseudomonas fluorescens in inhibition of Rhizoctonia solani, the rice sheath blight pathogen[J]. Microbiological Research,2004,159(1):73-81.
[9] ASAKA O,SHODA M. Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB 14[J]. Applied Microbiology,1996,62(11):4081-4085.
[10] RYU C M, KIM J W, CHOI O, et al. Nature of a root-associated Paenibacillus polymyxa from field-grown winter barley in Korea[J]. Journal of Microbiology and Biotechnology, 2005, 15(5):984-991.
[11] BAKI A A, ANDERSON J D. Vigour determination determination in soyabean seed by multiple criteria[J]. Crop Science,1973,31:630-633.
[12] 任爭光,張志勇,魏艷敏.芽孢桿菌防治園藝植物病害的研究進展[J].中國生物防治,2006(Z1):194-198.
[13] 凌世海.淺談我國農藥水分散粒劑的開發[J].安徽化工,2004,30(1):2-4.
[3] STRASHNOV Y, ELAD Y, SIVAN A, et al. Control of Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai [J]. Crop Protection,1985,4:359-364.
[4] SECOR G A, GUDMESTAD NC. Managing fungal diseases of potato[J]. Canadian Journal of Plant Pathology, 1999, 21: 213-221.
[5] Parry D W. Diseases of potato[A]. Plant pathology in agriculture [M]. UK: Cambridge University Press,1981.
[6] REITHNER B, SCHUHMACHER R, STOPPACHER N, et al. Signaling via the Trichoderma atroviride mitogen-activated protein kinase Tmk1 differentially affects mycoparasitism and plant protection[J]. Fungal Genetics and Biology:FG&B,2007,44(11): 1123-1133.
[7] HADAR Y, CHET I, HENIS Y. Biological control of Rhizoctonia solani damping-off with wheat bran culture of Trichoderma harzianum[J]. Phytopathology,1979,69:64-68.
[8] NAGARAJKUMAR M, BHASKARAN R, VELAZHAHAN R. Involvement of secondary metabolites and extracellular lytic enzymes produced by Pseudomonas fluorescens in inhibition of Rhizoctonia solani, the rice sheath blight pathogen[J]. Microbiological Research,2004,159(1):73-81.
[9] ASAKA O,SHODA M. Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB 14[J]. Applied Microbiology,1996,62(11):4081-4085.
[10] RYU C M, KIM J W, CHOI O, et al. Nature of a root-associated Paenibacillus polymyxa from field-grown winter barley in Korea[J]. Journal of Microbiology and Biotechnology, 2005, 15(5):984-991.
[11] BAKI A A, ANDERSON J D. Vigour determination determination in soyabean seed by multiple criteria[J]. Crop Science,1973,31:630-633.
[12] 任爭光,張志勇,魏艷敏.芽孢桿菌防治園藝植物病害的研究進展[J].中國生物防治,2006(Z1):194-198.
[13] 凌世海.淺談我國農藥水分散粒劑的開發[J].安徽化工,2004,30(1):2-4.
[3] STRASHNOV Y, ELAD Y, SIVAN A, et al. Control of Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai [J]. Crop Protection,1985,4:359-364.
[4] SECOR G A, GUDMESTAD NC. Managing fungal diseases of potato[J]. Canadian Journal of Plant Pathology, 1999, 21: 213-221.
[5] Parry D W. Diseases of potato[A]. Plant pathology in agriculture [M]. UK: Cambridge University Press,1981.
[6] REITHNER B, SCHUHMACHER R, STOPPACHER N, et al. Signaling via the Trichoderma atroviride mitogen-activated protein kinase Tmk1 differentially affects mycoparasitism and plant protection[J]. Fungal Genetics and Biology:FG&B,2007,44(11): 1123-1133.
[7] HADAR Y, CHET I, HENIS Y. Biological control of Rhizoctonia solani damping-off with wheat bran culture of Trichoderma harzianum[J]. Phytopathology,1979,69:64-68.
[8] NAGARAJKUMAR M, BHASKARAN R, VELAZHAHAN R. Involvement of secondary metabolites and extracellular lytic enzymes produced by Pseudomonas fluorescens in inhibition of Rhizoctonia solani, the rice sheath blight pathogen[J]. Microbiological Research,2004,159(1):73-81.
[9] ASAKA O,SHODA M. Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB 14[J]. Applied Microbiology,1996,62(11):4081-4085.
[10] RYU C M, KIM J W, CHOI O, et al. Nature of a root-associated Paenibacillus polymyxa from field-grown winter barley in Korea[J]. Journal of Microbiology and Biotechnology, 2005, 15(5):984-991.
[11] BAKI A A, ANDERSON J D. Vigour determination determination in soyabean seed by multiple criteria[J]. Crop Science,1973,31:630-633.
[12] 任爭光,張志勇,魏艷敏.芽孢桿菌防治園藝植物病害的研究進展[J].中國生物防治,2006(Z1):194-198.
[13] 凌世海.淺談我國農藥水分散粒劑的開發[J].安徽化工,2004,30(1):2-4.